Abstract
It has been found that tissues fixed for electron microscopy and dehydrated in acetone can be embedded in mixtures of n-butyl methacrylate and polyester resin. Activation with 1 per cent tert-butyl hydroperoxide followed by 12 to 48 hours at 60°C produces blocks that section well with glass knives. The ribbons are cleared of methacrylate by heat (200–250°C for 1 hour) and/or immersion in organic solvents (CCL4, acetone-ether). After removal of the methacrylate the residual polyester matrix provides thermostable and insoluble support for the tissue. Its insolubility permits staining by immersion of cleared preparations in organic solvents carrying heavy metal compounds in solution. Clearing by heat stabilizes section-grid relationships. The removal of volatile materials by clearing substantially reduces contamination of both specimen and microscope. Tissue fine structure is well preserved in these preparations.
Full Text
The Full Text of this article is available as a PDF (674.5 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- FARQUHAR M. G. Preparation of ultrathin tissue sections for electron microscopy; review and compilation of procedures. Lab Invest. 1956 Jul-Aug;5(4):317–337. [PubMed] [Google Scholar]
- FINCK H. Epoxy resins in electron microscopy. J Biophys Biochem Cytol. 1960 Feb;7:27–30. doi: 10.1083/jcb.7.1.27. [DOI] [PMC free article] [PubMed] [Google Scholar]
- GLAUERT A. M., GLAUERT R. H., ROGERS G. E. A new embedding medium for electron microscopy. Nature. 1956 Oct 13;178(4537):803–803. doi: 10.1038/178803a0. [DOI] [PubMed] [Google Scholar]
- KELLENBERGER E., RYTER A., SCHWAB W. L'utilisation d'un copolymère du groupe des polyesters comme matériel d'inclusion en ultramicrotomie. Experientia. 1956 Nov 15;12(11):421–422. doi: 10.1007/BF02157363. [DOI] [PubMed] [Google Scholar]
- LEDUC E., BERNHARD W. [Attempts at ultrastructural cytochemistry. Action on chromatin]. C R Hebd Seances Acad Sci. 1960 Apr 25;250:2948–2950. [PubMed] [Google Scholar]
- LUFT J. H. Improvements in epoxy resin embedding methods. J Biophys Biochem Cytol. 1961 Feb;9:409–414. doi: 10.1083/jcb.9.2.409. [DOI] [PMC free article] [PubMed] [Google Scholar]
- LUFT J. H. Permanganate; a new fixative for electron microscopy. J Biophys Biochem Cytol. 1956 Nov 25;2(6):799–802. doi: 10.1083/jcb.2.6.799. [DOI] [PMC free article] [PubMed] [Google Scholar]
- PALADE G. E. A study of fixation for electron microscopy. J Exp Med. 1952 Mar;95(3):285–298. doi: 10.1084/jem.95.3.285. [DOI] [PMC free article] [PubMed] [Google Scholar]
- PORTER K. R., MACHADO R. D. Studies on the endoplasmic reticulum. IV. Its form and distribution during mitosis in cells of onion root tip. J Biophys Biochem Cytol. 1960 Feb;7:167–180. doi: 10.1083/jcb.7.1.167. [DOI] [PMC free article] [PubMed] [Google Scholar]
- RYTER A., KELLENBERGER E. L'inclusion au polyester pour l'ultramicrotomie. J Ultrastruct Res. 1958 Dec;2(2):200–214. doi: 10.1016/s0022-5320(58)90018-2. [DOI] [PubMed] [Google Scholar]
- WATSON M. L. Reduction of heating artifacts in thin sections examined in the electron microscope. J Biophys Biochem Cytol. 1957 Nov 25;3(6):1017–1022. doi: 10.1083/jcb.3.6.1017. [DOI] [PMC free article] [PubMed] [Google Scholar]